| Quinoa (Chenopodium quinoa), a dicotyledon originally produced in South Africa, is Chenopodiaceae and Chenopodium. It boasts unique, rich and comprehensive nutritional value. This paper takes quinoa as the raw material to extract flavonoids. Then, isolation and purification is conducted to it. It explores quinoa flavonoids’ antioxidant effect, drosophila anti-aging molecular mechanism, thereby providing theoretical basis for applying quinoa flavonoids to healthcare foods. The main research results are as follows:(1) Quinoa flavonoids extraction process optimization. Through orthogonal experiment,the optimum extraction process parameters were:temperature at 90℃, liquor ratio at 1:10 (g/mL), refluxing time for 2 h and ethanol concentration at 90%. Under this condition, the extraction capacity of total flavonoids of quinoa was 3.861 mg/g.(2) Using macroporous resin purification quinoa flavonoids extracts.Comparison of four kinds of resins (HPD400, AB-8, D001, D101) for total flavonoids quinoa static and dynamic adsorption, desorption performance,one of the best is D101,on the purification process optimization,the results show that the optimal purification process of macroporous resin was:pH 2, flow velocity 2 BV/h,0.3 mg/mL flavonoids aqueous solution,8 BV water washing, acetone elution. The purified flavonoids adsorption rate under these conditions was 80.91%. The purity could be improved from 7.12% of the original extracts to 28.53%.(3) The total flavonoids of quinoa with D101 resin purification were used to perform Sephadex LH-20 purification process study. The optimum process condition was that the flow velocity was 1mL/min, and methanol aqueous solutions that were 30%,40%,60%,70%,90% and 100% were used to elute columns for 1h, respectively. Under this process condition, total flavonoids content was 81.57%, and monomeric flavonoids were separated. It was determined by HPLC that the purity was 93.63%.(4) Vc and BHT as control,The total flavonoids of quinoa before and after D101 resin purification were used to perform DPPH,·OH, H2O2 scavenging and metal ion chelation,the result presented dose-dependent relations in a certain range, which had significant in vitro antioxidant activity.(5) Compared with the control groups, the average lifetime, half-survival lifetime and maximum lifetime of male and female flies were prolonged significantly (P<0.05) through fruit fly survival experiment with the flavonoids dosage groups of 50μg/mL,200 μg/mL and 500 μg/m.The increased range was 1.25～28.68%. However, compared with control groups, the survival time of male and female fruit flies was shortened significantly (P<0.05) in 1 mg/mL and 2 mg/mL dosage groups, and the shortened range was 1.08～20.63%. In the in vivo experiment, the total SOD vitality, MnSOD vitality, CuZnSOD vitality and CAT vitality of fruit flies in 50μg/mL,200 μg/mL and 500 μg/mL dosage groups were determined, respectively. Except that MnSOD vitality was not improved significantly (P<0.05) in male dosage groups, compared with control groups, the maximum enzyme activity of the rest male and female related antioxidant enzyme was improved significantly (P<0.05). The increased range was 0.61～35.88%, where the maximum increase of CAT vitality in the 200 μg/mL dosage group of female flies was up to 35.88%. In addition, there was significant (P<0.05) positive correlation between certain enzyme activity and survival index.(6) For the experimental groups compared with control groups, the real-time fluorescence quantification PCR technical testing result showed that the quinoa flavonoids of 50 μg/mL,200 μg/mL and 500 μg/mL dosage groups up-regulated CuZnSOD, CAT, Egfr, S6K and MAPK genes of male and female flies significantly (P<0.05), and the relative expression level of MnSOD genes for female flies. Besides, the relative expression level of Mth genes was down-regulated significantly (P<0.05). There was a significant (P<0.05) positive correlation between the antioxidation related genetic expression level and enzyme activity。... |
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